Study of glioma infiltration in complex microenvironment on an automated microfluidic chip

Research Project

Project/Area Number	17J00362
Research Category	Grant-in-Aid for JSPS Fellows
Allocation Type	Single-year Grants
Section	国内
Research Field	Biomedical engineering/Biomaterial science and engineering
Research Institution	Okinawa Institute of Science and Technology Graduate University
Research Fellow	TSAI Hsieh Fu 沖縄科学技術大学院大学, 科学技術研究科, 特別研究員(DC1)
Project Period (FY)	2017-04-26 – 2020-03-31
Project Status	Granted (Fiscal Year 2019)
Budget Amount *help	¥3,100,000 (Direct Cost: ¥3,100,000) Fiscal Year 2019: ¥1,000,000 (Direct Cost: ¥1,000,000) Fiscal Year 2018: ¥1,000,000 (Direct Cost: ¥1,000,000) Fiscal Year 2017: ¥1,100,000 (Direct Cost: ¥1,100,000)
Keywords	neuron culture / deep learning / label-free segmentation / electrotaxis / temperature controller
Outline of Annual Research Achievements	1. A facile PDMS treatment protocol is developed to support the growth of the most fastidious cell type in microsystems. 2. A open source versatile platform is developed for lab on chip experiments. It is very easy to program and can connect with various sensors and outputs. 3. A modified deep learning platform DeepCell is used for label-free cell segmentation and tracking the glioma cell migration. 4. Influence of extracellular matrix for glioma cell adhesion and electrotaxis. I've started to analyze the influence of ECM proteins on glioma cell adhesion and electrotaxis.
Current Status of Research Progress	Current Status of Research Progress 2: Research has progressed on the whole more than it was originally planned. Reason I am proceeding as research as planned. A microfluidic chip is designed and used for collecting of glioma cell electrotaxis under various microenvironment conditions. An deep-learning based label free phase contrast cell segmentation platform is also developed for resolving a major data analysis bottleneck for cell migration studies. An opensource temperature controller is also developed to fulfill partially the automated chip platform. Based on the advances in the first year, for the next year, i can move on to collect more cell electrotaxis data and investigate further how microenvironment affects the electrotaxis.
Strategy for Future Research Activity	The primary work for next year is to explore more on the electrotaxis of glioma cells when there are various coexisting chemical ligands in the microenvironment. Secondly, I have tested building a microfluidic chip that creates orthogonal chemical and voltage gradients. However, the stability of the gradients is prone to system perturbance (i.g. syringe pump instability, chip fabrication inperfections). Redesigning a microfluidic chip with stable gradient generation will be a key step in next year. Thirdly, the effective charge of native proteins is a grossly overlooked characteristic that is very important in our configuration. I intend to examine the effective charge of proteins by capillary electrophoresis examination this year.